Microspore-derived cell suspension cultures of oilseed rape as a system for studying gene expression

Shi, Yongzhong; Xu, Gaosi; Warrington, Timothy B.; Murdoch, Gordon K.; Kazala, E. Chris; Snyder, Crystal L.; Weselake, Randall J.

doi:10.1007/s11240-007-9313-5

Cited by 12 publications

(7 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The truncated version of BnDGAT1 cDNA probably represents a mutation produced through a duplication event that may have occurred over many years of sub-culturing (Nykiforuk et al 2002). Recent analysis of the nuclear DNA content of this culture by flow cytometry has shown an increase in DNA content relative to previous values for B. napus suggesting that chromosomal changes in the cells occurred over time resulting in aneuploidy (Shi et al 2008). The antisense construct (pD3) we used in this study is expected to suppress both long and short form of BnDGAT1 because of their extensive nucleotide identity (99%), but not the BnDGAT2 which presumably has no identity with our antisense construct.…”

Section: Resultsmentioning

confidence: 86%

Antisense suppression of type 1 diacylglycerol acyltransferase adversely affects plant development in Brassica napus

Lock¹,

Snyder

Zhu³

et al. 2009

Physiologia Plantarum

Self Cite

View full text Add to dashboard Cite

Diacylglycerol acyltransferase (DGAT) catalyzes the acyl-coenzyme A (CoA) dependent acylation of sn-1,2-diacylglycerol to form triacylglycerol in the terminal step of seed oil formation. Previous work has suggested that the level of DGAT activity may have a substantial effect on the flow of carbon into triacylglycerol, implying that the enzyme may represent a promising target for seed oil modification through biotechnological approaches. In the current study, Brassica napus DH12075 was transformed with an antisense type 1 DGAT construct, resulting in a reduction in DGAT1 gene expression, total DGAT activity and seed oil content. In addition, reduced seed yield and germination rates were observed along with severe developmental abnormalities. These data suggest that in addition to its critical role in seed oil formation, DGAT1 enzyme may also be important for normal seed development in B. napus, although the underlying mechanism(s) remain to be determined.

show abstract

Section: Resultsmentioning

confidence: 86%

Antisense suppression of type 1 diacylglycerol acyltransferase adversely affects plant development in Brassica napus

Lock¹,

Snyder

Zhu³

et al. 2009

Physiologia Plantarum

Self Cite

View full text Add to dashboard Cite

show abstract

“…B. napus cv Jet Neuf suspension cell cultures were grown in NLN medium, as previously described (36). Explicit details can be found in SI Materials and Methods.…”

Section: Methodsmentioning

confidence: 99%

Feedback regulation of plastidic acetyl-CoA carboxylase by 18:1-acyl carrier protein in Brassica napus

André

Haslam

Shanklin

2012

Proc. Natl. Acad. Sci. U.S.A.

140

115

View full text Add to dashboard Cite

Plant seed oil represents a major renewable source of reduced carbon, but little is known about the biochemical regulation of its synthesis. The goal of this research was to identify potential feedback regulation of fatty acid biosynthesis in Brassica napus embryo-derived cell cultures and to characterize both the feedback signals and enzymatic targets of the inhibition. Fatty acids delivered via Tween esters rapidly reduced the rate of fatty acid synthesis in a dose-dependent and reversible manner, demonstrating the existence of feedback inhibition in an oil-accumulating tissue. Tween feeding did not affect fatty acid elongation in the cytosol or the incorporation of radiolabeled malonate into nascent fatty acids, which together pinpoint plastidic acetyl-CoA carboxylase (ACCase) as the enzymatic target of feedback inhibition. To identify the signal responsible for feedback, a variety of Tween esters were tested for their effects on the rate of fatty acid synthesis. Maximum inhibition was achieved upon feeding oleic acid (18:1) Tween esters that resulted in the intracellular accumulation of 18:1 free fatty acid, 18:1-CoA, and 18:1-acyl-carrier protein (ACP). Direct, saturable inhibition of ACCase enzyme activity was observed in culture extracts and in extracts of developing canola seeds supplemented with 18:1-ACP at physiological concentrations. A mechanism for feedback inhibition is proposed in which reduced demand for de novo fatty acids results in the accumulation of 18:1-ACP, which directly inhibits plastidic ACCase, leading to reduced fatty acid synthesis. Defining this mechanism presents an opportunity for mitigating feedback inhibition of fatty acid synthesis in crop plants to increase oil yield.

show abstract

“…B. napus (L. cv Jet Neuf) microsporederived (BnaMD) cell suspension cultures were developed in 1984 and maintained in various laboratories. These cultures have been used for many scientific studies in the biochemistry and molecular biology of storage lipid biosynthesis, the enzymology of oilseed carbohydrate metabolism and abiotic stress [3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20][21]. We are particularly interested in examining molecular strategies for increasing the oil content of plant biomass [22].…”

Section: Introductionmentioning

confidence: 99%

A Rapid Nile Red Fluorescence‐Based Method for Triacylglycerol Content in Microspore‐Derived Cell Suspension Cultures of Brassica napus

Gao

Chen

Weselake

2014

Lipids

Self Cite

View full text Add to dashboard Cite

Brassica napus (L. cv Jet Neuf) microspore-derived (BnaMD) cell suspension cultures were used to study the biochemistry and molecular biology of plant lipid metabolism. The cell suspension cultures may also be a potentially useful trait-testing tool for further improvement of oilseed rape, the world's third largest vegetable oil crop. Nile red fluorescence methods have been used to assess neutral lipid accumulation in microorganisms and mammalian oocytes but have not been established for plant cell suspension cultures. In this study, a Nile red method was established and optimized for triacylglycerol (TAG) content analysis in BnaMD cell suspension cultures. Nile red fluorescence was easily determined in 96-well plates using a 490 nm excitation wavelength and a 595 nm emission wavelength without any significant interference from other cellular components. The lipid content of the cell suspension cultures at different growth stages was determined using a conventional lipid analysis method and the Nile red fluorescence method. There was a strong positive correlation between the Nile red fluorescence and TAG content in the cell suspension cultures. Therefore, the TAG content of BnaMD cell suspension cultures can be rapidly and robustly determined using a Nile red fluorescence-based method that can potentially be adapted for application to other plant cell suspension cultures.

show abstract

Microspore-derived cell suspension cultures of oilseed rape as a system for studying gene expression

Cited by 12 publications

References 42 publications

Antisense suppression of type 1 diacylglycerol acyltransferase adversely affects plant development in Brassica napus

Antisense suppression of type 1 diacylglycerol acyltransferase adversely affects plant development in Brassica napus

Feedback regulation of plastidic acetyl-CoA carboxylase by 18:1-acyl carrier protein in Brassica napus

A Rapid Nile Red Fluorescence‐Based Method for Triacylglycerol Content in Microspore‐Derived Cell Suspension Cultures of Brassica napus

Contact Info

Product

Resources

About